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. 2022 Apr;414(10):3243-3255.
doi: 10.1007/s00216-021-03847-x. Epub 2021 Dec 22.

An integrated device for fast and sensitive immunosuppressant detection

Sara Tombelli  1 , Cosimo Trono  2 , Simone Berneschi  1 , Chiara Berrettoni  1 , Ambra Giannetti  1 , Romeo Bernini  3 , Gianluca Persichetti  3 , Genni Testa  3 , Guillermo Orellana  4 , Francesca Salis  4 , Susanne Weber  5 , Peter B Luppa  5 , Giampiero Porro  6 , Giovanna Quarto  6 , Markus Schubert  7 , Marcel Berner  8 , Paulo P Freitas  9 , Susana Cardoso  9 , Fernando Franco  9 , Vânia Silverio  9 , Maria Lopez-Martinez  9 , Urs Hilbig  10 , Kathrin Freudenberger  10 , Günter Gauglitz  10 , Holger Becker  11 , Claudia Gärtner  11 , Mark T O'Connell  12 , Francesco Baldini  1
Affiliations

An integrated device for fast and sensitive immunosuppressant detection

Sara Tombelli et al. Anal Bioanal Chem. 2022 Apr.

Abstract

The present paper describes a compact point of care (POC) optical device for therapeutic drug monitoring (TDM). The core of the device is a disposable plastic chip where an immunoassay for the determination of immunosuppressants takes place. The chip is designed in order to have ten parallel microchannels allowing the simultaneous detection of more than one analyte with replicate measurements. The device is equipped with a microfluidic system, which provides sample mixing with the necessary chemicals and pumping samples, reagents and buffers into the measurement chip, and with integrated thin film amorphous silicon photodiodes for the fluorescence detection. Submicrometric fluorescent magnetic particles are used as support in the immunoassay in order to improve the efficiency of the assay. In particular, the magnetic feature is used to concentrate the antibody onto the sensing layer leading to a much faster implementation of the assay, while the fluorescent feature is used to increase the optical signal leading to a larger optical dynamic change and consequently a better sensitivity and a lower limit of detection. The design and development of the whole integrated optical device are here illustrated. In addition, detection of mycophenolic acid and cyclosporine A in spiked solutions and in microdialysate samples from patient blood with the implemented device are reported.

Keywords: Cyclosporine A; Fluorescence; Immunosuppressant; Mycophenolic acid; POCT.

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Conflict of interest statement

The author GG is an editor of the journal, but as such was not involved in the peer review of this article. The other authors declare no conflict of interest.

Figures

Fig. 1
Fig. 1
Exploded view of the core of the POCT system
Fig. 2
Fig. 2
Scheme of the different steps of the on-chip immunoassay
Fig. 3
Fig. 3
a 3D scheme of the microfluidic chip. b The three components of the optical chip. From left to right: double-sided adhesive tape with laser-cut microchannels, molded Zeonex® top part with integrated fluidic ports and Zeonor® bottom foil. c Magnified view of the cross section of the microfluidic chip depicting two of the channels (the yellow lines represent the optical path of the fluorescence rays)
Fig. 4
Fig. 4
Schematic view of the fluidic system based on Fluigent® pressure-driven microfluidics pumps
Fig. 5
Fig. 5
Actual optical integrated device interfaced to the tablet PC
Fig. 6
Fig. 6
Evaluation of the crosstalk between the microfluidic/optical detection channels (ch). Light gray bars: normalized fluorescence from all the channels filled with PBS; dark gray bars: fluorescence from the FMP attached to the bottom surface (Zeonor®) of ch7 microfluidic channel
Fig. 7
Fig. 7
Detected optical signals from four different channels filled with PBS buffer after successively loading the chip inside the optical device five times. The standard deviations (σ) of the five different acquired signals are indicated
Fig. 8
Fig. 8
a Calibration curve for CsA obtained with the optical device using FMPs coated with anti-CsA antibody and CsA diluted in 20% Lipofundin. b Calibration curve for MPA obtained with the optical device by using FMPs coated with anti-MPA antibody and MPA diluted in 20% Lipofundin
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References

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